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The main objective of this study was the detection, identification and expression analysis of genes associated with blastocyst formation in order to help us better understand this process. This information could lead to improvements of\n                      <jats:italic>in vitro<\/jats:italic>\n                      embryo production procedures.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      A subtractive cDNA library was constructed enriched for transcripts preferentially expressed at the blastocyst stage compared to the 2-cell and 8-cell stage. Sequence information was obtained for 65 randomly selected clones. The RNA expression levels of 12 candidate genes were determined throughout 3 stages of preimplantation embryo development (2-cell, 8-cell and blastocyst) and compared with the RNA expression levels of\n                      <jats:italic>in vivo<\/jats:italic>\n                      \"golden standard\" embryos using real-time PCR. The RNA expression profiles of 9 (75%) transcripts (\n                      <jats:italic>KRT18<\/jats:italic>\n                      ,\n                      <jats:italic>FN1<\/jats:italic>\n                      ,\n                      <jats:italic>MYL6<\/jats:italic>\n                      ,\n                      <jats:italic>ATP1B3<\/jats:italic>\n                      ,\n                      <jats:italic>FTH1<\/jats:italic>\n                      ,\n                      <jats:italic>HINT1<\/jats:italic>\n                      ,\n                      <jats:italic>SLC25A5<\/jats:italic>\n                      ,\n                      <jats:italic>ATP6V0B<\/jats:italic>\n                      ,\n                      <jats:italic>RPL10<\/jats:italic>\n                      ) were in agreement with the subtractive cDNA cloning approach, whereas for the remaining 3 (25%) (\n                      <jats:italic>ACTN1<\/jats:italic>\n                      ,\n                      <jats:italic>COPE<\/jats:italic>\n                      ,\n                      <jats:italic>EEF1A1<\/jats:italic>\n                      ) the RNA expression level was equal or even higher at the earlier developmental stages compared to the blastocyst stage. Moreover, significant differences in RNA expression levels were observed between\n                      <jats:italic>in vitro<\/jats:italic>\n                      and\n                      <jats:italic>in vivo<\/jats:italic>\n                      produced embryos. By immunofluorescent labelling, the protein expression of KRT18, FN1 and MYL6 was determined throughout bovine preimplantation embryo development and showed the same pattern as the RNA expression analyses.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      By subtractive cDNA cloning, candidate genes involved in blastocyst formation were identified. For several candidate genes, important differences in gene expression were observed between\n                      <jats:italic>in vivo<\/jats:italic>\n                      and\n                      <jats:italic>in vitro<\/jats:italic>\n                      produced embryos, reflecting the influence of the\n                      <jats:italic>in vitro<\/jats:italic>\n                      culture system on the embryonic gene expression. Both RNA and protein expression analysis demonstrated that\n                      <jats:italic>KRT18<\/jats:italic>\n                      ,\n                      <jats:italic>FN1<\/jats:italic>\n                      and\n                      <jats:italic>MYL6<\/jats:italic>\n                      are differentially expressed during preimplantation embryo development and those genes can be considered as markers for bovine blastocyst formation.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/1471-213x-7-64","type":"journal-article","created":{"date-parts":[[2007,6,8]],"date-time":"2007-06-08T14:13:56Z","timestamp":1181312036000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":54,"title":["Identification and expression analysis of genes associated with bovine blastocyst formation"],"prefix":"10.1186","volume":"7","author":[{"given":"Karen","family":"Goossens","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ann","family":"Van Soom","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mario","family":"Van Poucke","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leen","family":"Vandaele","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jo","family":"Vandesompele","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alex","family":"Van Zeveren","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luc J","family":"Peelman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2007,6,8]]},"reference":[{"key":"206_CR1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1002\/bdrc.20035","volume":"75","author":"C Wrenzycki","year":"2005","unstructured":"Wrenzycki C, Herrmann D, Lucas-Hahn A, Gebert C, Korsawe K, Lemme E, Carnwath JW, Niemann H: Epigenetic reprogramming throughout preimplantation development and consequences for assisted reproductive technologies. 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